European Journal of Medicinal Chemistry 46 (2011) 526e534

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European Journal of Medicinal Chemistry

journal homepage: http://www.elsevier.com/locate/ejmech

Original article

Synthesis and anti-inflammatory evaluation of some new acyl-hydrazones

bearing 2-aryl-thiazole
Cristina Mariana Moldovan a, *, Ovidiu Oniga a, Alina Pârvu b, Brînduşa Tiperciuc a, Philippe Verite c,
u d, Ovidiu Crişan e, Marius Bojiţa
Adrian Pîrna  f, Raluca Pop g
a
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Medicine and Pharmacy, 41 Victor Babes Street, RO-400010 Cluj-Napoca, Romania
b
Department of Physiology, Faculty of Medicine, University of Medicine and Pharmacy, Victor Babes Street, Cluj-Napoca, Romania
c
Department of Analytical Chemistry, Faculty of Pharmacy, University of Medicine and Pharmacy, 22 Gambetta Boulevard, F-76183 Rouen Cedex, France
d
National Institute for Research and Development of Isotopic and Molecular Technologies, RO-400293 Cluj-Napoca, Romania
e
Department of Organic Chemistry, Faculty of Pharmacy, University of Medicine and Pharmacy, 41 Victor Babes Street, RO-400010 Cluj-Napoca, Romania
f
Department of Drug Analysis, Faculty of Pharmacy, University of Medicine and Pharmacy, 6 Louis Pasteur Street, Cluj-Napoca, Romania
g
National Institute for Research and Development for Electrochemistry and Condensed Matter, 144 Aurel Paunescu-Podeanu Street, RO-300569, Timişoara, Romania

a r t i c l e i n f o a b s t r a c t

Article history: This work describes recent results from our research program aiming at the synthesis and evaluation of
Received 25 May 2010 new compounds acting as potential anti-inflammatory drugs. A series of novel acyl-hydrazones bearing
Received in revised form 2-aryl-thiazole moiety were synthesized by the condensation between derivatives of 4-[2-(4-methyl-2-
9 November 2010
phenyl-thiazole-5-yl)-2-oxo-ethoxy]-benzaldehyde and 2, 3 or 4-(2-aryl-thiazol-4-ylmethoxy)-benzal-
Accepted 21 November 2010
Available online 1 December 2010
dehyde, respectively and different carboxylic acid hydrazides. The structures of newly synthesized
compounds were established by the combined use of IR, 1H NMR, mass spectral data and elemental
analysis. These compounds were tested in vivo for their anti-inflammatory activity, in an acute experi-
Keywords:
Acyl-hydrazone
mental inflammation. The acute phase bone marrow response, phagocytes’ activity and NO synthesis
2-Aryl-thiazole were evaluated. Compounds 10, 15, 17, 18 and 22 reduced the absolute leukocytes count due to the lower
Anti-inflammatory activity neutrophils percentage. Phagocitary index was decreased by all the compounds. Seven of them reduced
the phagocitary activity. Five compounds inhibited NO synthesis, 3, 4, 16 and 22 stronger than Melox-
icam, the anti-inflammatory reference drug.
Ó 2010 Elsevier Masson SAS. All rights reserved.

1. Introduction acute and chronic inflammation by producing nitric oxide as

Rheumatic diseases are the most prevalent causes of disability
in European countries and non-steroidal anti-inflammatory drugs
(NSAIDs) are still the most commonly used remedies. Chronic use
may cause several serious adverse effects, the most important one
being gastric injury and renal complications. GI damage from
NSAID is generally attributed to two factors: local irritation by the
direct contact of the free carboxylic acid (eCOOH) moiety of NSAID
with GI mucosal cells (topical effect) and decreased tissue prosta-
glandin production in tissues [1].
The development of new non-steroidal anti-inflammatory drugs
follows different strategies: selective COX-2 inhibition or the inhi-
bition of inducible nitric oxide synthase (iNOS). iNOS contributes to
* Corresponding author. Present address: 2B Calea Floresti Street, A1B Building,
Apartment 25, Cluj-Napoca, Romania. Tel.: þ40 745 264393; fax: þ40 264 450529.
E-mail addresses: cris.moldovan@yahoo.com, cmoldovan@umfcluj.ro
(C.M. Moldovan).
a cytotoxic inflammatory mediator [2]. Synthetic approaches have
been taken with the aim of improving safety profile and in turn
therapeutic window of NSAIDs. Several studies have described the
derivatization of the carboxylate function of representative NSAID
with less acidic azoles: thiazole [3e8], oxadiazole [9e12], triazole,
thiadiazole [2], etc. which resulted in an increased anti-inflam-
matory activity with reduced ulcerogenicity.
In our attempt to synthesize new, safer and potent agents for
treatment of inflammatory diseases, we used 2-aryl-thiazole
moiety. This heterocyclic system has found application in drug
development for the treatment of inflammation [13]. Fentiazac
(2-phenyl-thiazole), Meloxicam (2-methyl-thiazole), Fanetizole
(2-amino-thiazole) [14e16] are some examples of thiazole bearing
anti-inflammatory products.
A number of hydrazone derivatives have been claimed to
possess anti-inflammatory activity [2,17e21]. Several studies pre-
sented the broad range of biological activities, including anti-
inflammatory, for compounds bearing chromone moiety [22].

0223-5234/$ e see front matter Ó 2010 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2010.11.032
 C.M. Moldovan et al. / European Journal of Medicinal Chemistry 46 (2011) 526e534
With the aim of obtaining new anti-inflammatory agents, we
designed a series of new acyl-hydrazones bearing 2-aryl-thiazole
scaffold. The compounds were evaluated for their effect on acute
inflammation.
2. Chemistry
The conversion of the aldehydes 1, 5e9 (Fig. 1) to the corre-
sponding acyl-hydrazones 3, 4, 10e18 (Scheme 1), 20e22 (Scheme
2) was readily accomplished by the reaction with the respective
carboxylic acid hydrazides, in refluxing acetic acid 50%.
The benzaldehyde derivatives 1 and 5e9 can be synthesized as
described in our previous paper by the etherification of a halogenated
component with 2, 3 or 4-hydroxy-benzaldehyde [23], and the
hydrazides 2a, b and 19 were prepared according to the literature by
refluxing thiazolyl acetic esters with hydrazine hydrate. The purity of
the compounds was demonstrated by TLC. All the new compounds
3, 4,10e18, 20e22 were characterized by m.p., elemental analysis and
spectroscopic data (IR, 1H NMR, 13C NMR, MS).
3. Pharmacology
Our study investigated the effects of the newly synthesized
compounds in an acute experimental inflammation, induced by the
i.m. injection of turpentine oil. Their anti-inflammatory capacity was
assessed by evaluating the acute phase bone marrow response,
phagocytes’ activity and NO synthesis. The acute phase bone marrow
response was measured by absolute leukocytes count and leukocytes
count expressed as percentage. Phagocytic activity was assessed with
the in vitro phagocytosis test by calculating two parameters: the
phagocytic index (PI) (PI% ¼ phagocytes with at least one phagocyted
germ from 200 leukocytes counted) and the phagocytic activity (PA)
(PA ¼ number of germs phagocyted by 100 leukocytes) [24e28]. NO
synthesis was evaluated measuring nitrates/nitrites concentration
[29,30]. The effects of new compounds were compared with those
from the inflammation group, and with those from the group treated
with Meloxicam as a reference NSAID. A negative control group of
healthy rats without any treatment was also used.
4. Results and discussion
4.1. Chemistry
The structures of all new compounds were confirmed by their
spectral (IR, 1H NMR, 13C NMR and mass) and elemental analytical
data. The IR spectra exhibited characteristic absorption band at
1675e1649 cm1 and 3446e3161 cm1 due to eCOe and eNH
stretching. Each of the new acyl-hydrazones displays a strong and
sharp band in the region 1555e1508 cm1 ascribed to n(C]N) of
azomethine group, providing confirmatory evidence for the
Fig. 1. Aldehydes 1 and 5e9.
527
condensation reaction between aldehyde group and hydrazide. The
1
H NMR spectra showed characteristic singlets due to NH and N]
CH proton at d 12.1e11 ppm and d 8.41e8.198 ppm, respectively.
The C(5)eH in thiazole appeared as one singlet or two singlet peaks
(compounds 10e18) at d 7.622e7.422 ppm, the protons from the
two methylene groups appeared as two singlet signals at
d 5.4e4.17 ppm and those from methyl group appeared at
d 2.785e2.34 ppm as singlet peak or two singlet peaks (compound
3). Other protons appeared in the aromatic regions. 13C NMR data
also supported the structures of acyl-hydrazones.
4.1.1. The structure of the compounds
The resulting double bond eHC]Ne following the condensation
reaction, contributes to the formation of geometrical isomers Z and E.
One distinctive feature to be remarked is the amide proton resonance
which is well downfield of all other peaks. Thus, the 1H NMR spectra
indicated the chemical shift of the NH proton at d 12.1e11 ppm in the
form of a singlet peak of an intense signal, assigned to the NH proton
of E isomer, accompanied by a weak singlet signal, assigned to the NH
proton of Z isomer, more deshielded.
The proton signals consistent with isomers Z and E in liquid state
were in agreement with ROESY spectra. These showed a stronger
interaction between the proton of NH group and that of eCH], both
protons with higher chemical shift (Z isomer), compared against the
interaction between the same two protons (NH and eCH]N) with
lower chemical shift, belonging to the E isomer. The formation of
both isomers is thermodynamically controlled, fact reflected in NMR
spectra, as differences in intensities of NH proton signals, corre-
sponding to both isomers. The NMR spectra showed a difference
between the intensities of the two peaks and the predominant
formation of one of the isomers. This one must have lower minimum
energy stabilization. HyperChem calculation [31], based on the
Polak-Ribiere (conjugate gradient) indicated for compound 10, for
example, a minimum energy stabilization of E isomer of 58.1345
kcal/mol, with a 0.08947 kcal/A*mol RMS gradient, whereas for
Z isomer the energy was 62.2176 kcal/mol, with a 0.09077 kcal/
A*mol RMS gradient. For all the other compounds, the minimum
energy stabilization of E isomer was lower than of Z isomer. The ab
initio calculation using Gaussian 03 [32] was possible after the
optimization of the geometry for the two isomers. The calculation
with this method indicated a total energy for E isomer(2262.493
a.u.), whereas for Z isomer (2262.472 a.u.) and a relative energy of
12.79 kcal/mol, comparing to the energy of E isomer, considered 0.
This indicates that E isomer is more stable.
Considering these observations, we can say that the two isomers
may coexist, in liquid state, but the E isomer is formed predomi-
nantly, this being in agreement with its higher stability and with
the literature data [33e35].
Hydrazones display prototropic amido-imido tautomerism. The
possibility of this type of tautomerism involving a hydroxyl group
was excluded for all the compounds, by the evident absence of typical
528 C.M. Moldovan et al. / European Journal of Medicinal Chemistry 46 (2011) 526e534

Scheme 1. Synthesis of acyl-hydrazones 3, 4 and 10e18.

signals of an OH group in IR and 1H NMR spectra. The IR absorption of
the hydrazone n(NH) group at 3446e3161 cm1, together with
a strong band at about 1675e1649 cm1 for the n(C]O) group,
confirms the amido tautomeric form in solid state.
4.2. Anti-inflammatory activity
4.2.1. Acute phase bone marrow response
Acute phase bone marrow response, as a part of the systemic
acute phase response, is induced by the early production of pro-
inflammatory cytokines IL-1, IL-6 and TNF-a, resulting leukocytosis
and neutrophilia. After 24 h from turpentine oil administration, it
was found that the synthesized compounds 10, 11, 15e18, 20 and 22
significantly reduced absolute leukocytes count (p < 0.001), 4 and
21 caused an important increase (p < 0.01) and 13 and 14 had no
significant influence (p > 0.05). Comparing the same results with
those from the Meloxicam group, we noticed that five compounds
(10, 11, 16e18) had a stronger inhibitory activity (p < 0.01) (Table 1).
Absolute leukocytes count reduction was positively correlated with
NO synthesis for substances 11, 15, 16 (r ¼ 0.7), with PI for
substances 17, 22 (r ¼ 0.7), and for PA with substance 18 (r ¼ 0.7).
Analyzing neutrophils percentage, we found a very significant
reduction in groups 3, 10, 13, 17, 18 and 22 (p < 0.001), a significant
decrease in groups 14 and 15 (p < 0.01) and a no significant change

Scheme 2. Synthesis of acyl-hydrazones 20e22.

 C.M. Moldovan et al. / European Journal of Medicinal Chemistry 46 (2011) 526e534 529

Table 1
Effects of the compounds on the acute medullar response.

Compound Leukocytesa (no./mm3) Neutrophilesa (%) Monocytesa (%) Lymphocytesa (%)

Control 4952.7  436.9 55.6  1.32 7.6  0.45 37  2.2
Inflammation 9280  412.79 73.43  1.9 2.28  0.76 27.71  4.53
Meloxicam 7660  422.37 66.57  4.27 2  1.15 32.28  4.23
3 8742.85  390.97 66  1.63 2.43  0.79 32.14  1.34
4 10200  556.77 78  1.15 2  0 18  4.16
10 3271.43  393.28 52.57  2.5 1.86  0.69 21.43  3.41
11 5664.28  355.57 82.28  2.93 2  0.58 20.71  3.68
12 8620  488.98 73.29  3.77 2.86  1.07 19.71  4.23
13 9540  333.46 61.71  1.8 2.4  0.84 32.28  3.35
14 9300  310.91 67.71  3.9 2.86  1.07 29.14  4.88
15 7837.14  203.11 68.85  2.79 2.57  0.79 24.71  3.77
16 6914.28  279.45 70.86  3.97 2.86  1.07 23.43  4.28
17 7314.28  481.07 68.28  2.43 2.71  0.95 28.57  2.7
18 7402.85  479.78 63.14  3.44 3.14  1.07 32  4.32
20 8228.57  275.16 76  3.83 3.71  0.75 23.57  4.47
21 12100  314.9 71.71  1.8 2.6  0.97 24  2.83
22 8000  273.86 57.43  2.76 3.28  0.95 39.86  2.48
a
Values are expressed as mean of three experiments  standard deviation.

in 12, 16 and 21. Two of the new molecules, 10 and 22, showed
a more potent inhibitory activity on neutrophils than Meloxicam
(p < 0.001) (Table 1). Neutrophils count expressed as percentage
was positively correlated with NO synthesis for substances 11, 17, 20
(r ¼ 0.7e0.8), with PI for substance 11 (r ¼ 0.7) and with substances
18, 21 for PA (r ¼ 0.8).
Compounds 10, 17, 18 and 22 showed an important reduction of
total leukocytes count by reducing neutrophils’ percentage.
Analyzing the chemical structures of these compounds, it can be
observed that the substitution of phenyl from position 2 of thiazole
with a brome atom in 4, had a good influence on the acute phase
bone marrow response (R ¼ Br for compounds 17, 18 and 22).
On the basis of these data, it could be argued that the
compounds with significant inhibitory activity observed in the
reduction of acute phase bone marrow response, have systemic
anti-inflammatory effects.
4.2.2. Phagocytosis test
Phagocytosis is part of the cellular acute phase response from
acute inflammations. There are two professional phagocytes:
polymorphonuclear (PMN) leukocytes and mononuclear phago-
cytes. The PMN cells are released from the bone marrow as mature
cells, which circulate in the blood before migrating into the tissues
where they perform their effector functions for 1 or 2 days. In
contrast, mononuclear phagocytes emerge from the marrow as
immature cells monocytes, circulate in the blood and then enter
tissues and organs of the body where they mature into macro-
phages. After recognizing the targets, phagocytic cells are activated
to ingesting them and destroying with reactive oxidants and
hydrolytic enzymes.
In our study, we have investigated phagocytic activity of blood
phagocytes for the evaluation of immunomodulating activity of the
synthesized agents. All the tested compounds caused a very
significant reduction of the PI (p < 0.001). Compared to Meloxicam,
just seven compounds (3, 4, 11, 13, 15, 20 and 22) had a stronger
inhibitory activity on PI (p < 0.001) (Fig. 2). As for the phagocytic
activity, it was observed that this parameter was very significantly
(p < 0.001) reduced by the compounds 3, 11, 13, 15, 16, 20 and 22.
Only four compounds (3, 11, 20 and 22) were more powerful
inhibitors of PA than Meloxicam (p < 0.05 for 3 and p < < 0.001 for
11, 20 and 22) (Fig. 3). PI reduction was positively correlated with
NO synthesis for substances 13, 15, 18, 22 (r ¼ 0.7), and with PA for
substance 11 (r ¼ 0.7).
Compounds 3, 11, 13, 15, 16, 20 and 22 had a significant influence
on the phagocytosis process, reducing PI and PA, too. These results
indicated that the presence of CF3 group in position meta of phenyl
determined an important decrease of phagocytary parameters
(X ¼ F for the compounds 11, 13, 15 and 16). Also, it should be
noticed the influence of the chromone group for the compounds 20
and 22.
4.2.3. Nitrites and nitrates concentration
In acute inflammation there is a significant increase of NO
synthesis due to the expression of iNOS. This will raise serum
nitrates/nitrites concentration, as side metabolites of NO. Melox-
icam reduced significantly NO synthesis (p < 0.001) (Fig. 4). For the

70

60

50

40
PI%

30

20

10

0
C I M 3 4 10 11 12 13 14 15 16 17 18 20 21 22

Fig. 2. The effect of the synthesized compounds on PI.

530 C.M. Moldovan et al. / European Journal of Medicinal Chemistry 46 (2011) 526e534
Fig. 3. The effect of the synthesized compounds on PA.
Fig. 4. The effect of the compounds on nitrates/nitrites amount.
compounds 10e12, 14, 18 and 20, the results showed a significant
increase (p < 0.05) of NO metabolites comparing with the Inflam-
mation group, while compounds 3, 4, 16, 21 and 22 reduced
significantly (p < 0.05) the amount of nitrates/nitrites and exhibi-
ted a stronger inhibitory activity (p < 0.01) of NO synthesis than
Meloxicam, excepting compound 21 (p > 0.05).
5. Conclusion
Various substituted 2-aryl-thiazole hydrazone derivatives were
synthesized and screened for their anti-inflammatory potential.
Compounds 10, 15, 17, 18 showed to have a good inhibitory effect on
the acute phase marrow response, by reducing the absolute
leukocytes count due to the lower neutrophils percentage.
Compound 10, which has 2-phenyl-thiazole and [2-(4-methyl-
phenyl)-4-methylen]-thiazole hydrazine moieties in its structure,
proved to be a more active inhibitor of the marrow acute phase
response than Meloxicam. From the results of in vitro phagocytosis
test we could conclude that all the newly synthesized compounds
reduced significantly PI, 3, 4, 11, 13, 15, 20 and 22 being more potent
inhibitors than Meloxicam. PA was significantly reduced by the
compounds 3, 11, 13, 15, 16, 20 and 22, from which 3, 11, 20 and 22
were more potent inhibitors than Meloxicam. The NO synthesis
was significantly reduced by 3, 4, 16, 21 and 22 and they all, except
of 21, showed a stronger inhibitory activity than Meloxicam.
6. Experimental section
6.1. Chemistry
Solvents were obtained from commercial sources; the reagents
were synthesized in our laboratory. Analytical thin layer chroma-
tography was carried out on precoated Silica Gel 60F254 sheets
using UV absorption for visualization. The melting points were
taken with two melting point meters, Electrothermal and MPM-H1
Schorpp and are uncorrected. FT-IR spectra were recorded on
a Nicolet 210 FT-IR spectrometer with a MCT detector and an Omnic
4.1b soft system, using potassium bromide. The 1H NMR and 13C
NMR spectra were recorded at room temperature on a Bruker
Avance NMR spectrometer operating at 500 MHz and 125 MHz,
respectively and were in accord with the assigned structures.
Chemical shift values were reported relative to tetramethylsilane
(TMS) as internal standard. The samples were prepared by dis-
solving the synthesized powder of the compounds in DMSO-d6
(dH ¼ 2.51 ppm, dC ¼ 40.02 ppm) as solvent and the spectra were
recorded using a single excitation pulse of 10.1 ms (1H NMR) and
8 ms (13C NMR), respectively. The two-dimensional NMR spectrum
at 500 MHz was obtained on standard Bruker software. The
conditions for ROESY phase-sensitive spectra via time proportional
phase incrementation (TPPI) were: spectral widths of 6.0 ppm in
both dimensions with a resolution of 0.7 and 1.4 Hz/point in f2 and
f1 respectively and a mixing time of 450 ms. The experiment was
performed using 4096 data points in f2 and 2048 in f1 increments
with 16 scans per t1 value and a relaxation period of 2 s. A sine
function (SSB ¼ 2) was applied in f1 and f2 before Fourier trans-
formation. GCeMS analyses were realized with an Agilent gas
chromatograph 6890 equipped with an apolar Macherey Nagel
Permabond SE 52 capillary column. Elemental analysis was regis-
tered with a Vario El CHNS instrument.
6.1.1. General procedure for the synthesis of acyl-hydrazones 3, 4,
10e18, 20e22
4-[2-(4-Methyl-2-phenyl-thiazol-5-yl)-2-oxo-ethoxy]-benzal-
dehyde 1 or 2, 3 or 4-(2-Aryl-thiazol-4-ylmethoxy)-benzaldehydes
5e9 (1 mmol) was dissolved in 10 ml of acetic acid 50%. Separately,
a suspension of acyl-hydrazide (1 mmol) in 10 ml acetic acid 50%
 C.M. Moldovan et al. / European Journal of Medicinal Chemistry 46 (2011) 526e534
was prepared. This suspension of hydrazide was added under
stirring to the solution of benzaldehyde. The reaction mixture was
stirred for 1.5 h. For improving the solubility of products in acetic
acid and the reaction, a small quantity of ethanol was added. After
1.5 h of refluxing, the reaction mixture was cooled off. The crude
product was filtered under reduced pressure, washed with water
on the filter and purified by crystallization from ethanol.
6.1.1.1. (2-p-Tolyl-thiazol-4-yl)-acetic acid {4-[2-(4-methyl-2-phenyl-
thiazol-5-yl)-2-oxo-ethoxy]-benzylidene}-hydrazide (3). Yield 53%.
Yellow powder, mp: 147e50  C. IR (KBr, cm1): 3219 (NH), 3064
(CHarom), 2921 and 2850 (CHaliph), 1674 (C]O), 1652 (C]O
Hydrazide), 1599 (C]C), 1554 (C]N). 1H NMR (DMSO-d6, 500 MHz,
ppm): d 11.95 (s, 1H, NH); 8.2 (s, 1H, eCH]); 7.96 (dd, 2H, Ar-H);
7.824e7.8 (m, 3H, Ar-H); 7.66 (dd, 2H, Ar-H); 7.511 (d, 2H, Ar-H);
7.448 (s, 1H, H-Thiazole); 7.302 (dd, 2H, Ar-H); 7.148 (dd, 2H, Ar-H);
5.292 (s, 2H, eCH2e); 4.183 (s, 2H, eCH2); 2.785 (s, 3H, eCH3),
2.35 (s, 3H, Ar-CH3). 13C NMR (DMSO-d6, 125 MHz, ppm): d 188.61
(C]O, ketone); 171.27 (C]O, hydrazide); 170.12 (C-2 Thiazole);
165.51 (C-4 Thiazole); 160.48 (C-2 Thiazole); 159.89 (]CeO, Ar);
153.39 (C-4 Thiazole); 151.61 (eCH]N); 143.21 (C, Ar); 140.01 (C,
Ar); 134.19 (C, Ar); 132.27 (C, Ar); 131.04 (C-5 Thiazole); 130.21 (2C,
CH, Ar); 128.84 (2C, CH, Ar); 128.54 (2C, CH, Ar); 128.02 (2C, CH,
Ar); 127.11 (2C, CH, Ar); 122.31 (CH, C-5 Thiazole); 118.78 (CH, Ar);
115.58 (2C, CH, Ar); 72.05 (CH2); 37.78 (CH2); 21.39 (CareCH3); 19.01
(C4thiazoleeCH3). Anal. Calcd. (%) for C31H26N4O3S2 (566.69): C,
65.70; H, 4.62; N, 9.89; S, 11.31. Found: C, 65.49; H, 4.61; N, 9.88; S,
11.28. MS (EI, 70 eV): m/z 567 (Mþ), 352 [M- C12H10NSO], 217
[C12H10NSO].
6.1.1.2. [2-(3-Trifluoromethyl-phenyl)-thiazol-4-yl]-acetic acid {4-[2-
(4-methyl-2-phenyl-thiazol-5-yl)-2-oxo-ethoxy]-benzylidene}-
hydrazide (4). Yield 80.64%. Yellow powder, mp: 167e9  C. IR (KBr,
cm1): 3161 (NH), 3060 (CHarom), 2919 and 2847 (CHaliph), 1670
(C]O), 1658 (C]O Hydrazide), 1587 (C]C), 1552 (C]N). 1H NMR
(DMSO-d6, 500 MHz, ppm): d 11.559 (s, 1H, NH); 8.26 (s, 1H,
eCH]); 8.18 (d, 2H, Ar-H); 7.994e7.952 (m, 3H, Ar-H); 7.81 (d, 2H,
Ar-H); 7.686e7.66 (m, 2H, Ar-H); 7.52 (s, 1H, H-Thiazole); 7.508 (d,
2H, Ar-H); 7.149 (dd, 2H, Ar-H); 5.286 (s, 2H, eCH2e); 4.22 (s, 2H,
eCH2e); 2.49 (s, 3H, eCH3). 13C NMR (DMSO-d6, 125 MHz, ppm):
d 188.65 (C]O, ketone); 171.19 (C]O, hydrazide); 170.2 (C-2
Thiazole); 165.04 (C-4 Thiazole); 160.43 (C-2 Thiazole); 159.5 (]
CeO, Ar); 152.16 (C-4 Thiazole); 146.92 (eCH]N); 143.2 (C, Ar);
134.38 (C, Ar); 132.49 (C, Ar); 132.13 (CH, Ar); 131.04 (CH, C-5
Thiazole); 130.53 (CH, Ar); 129.89 (2C, CH, Ar); 128.77 (C, Ar); 128.4
(CH, Ar); 128.01 (2C, CH, Ar); 127.14 (2C, CH, Ar); 125.41 (CF3);
123.24 (CH, Ar); 122.36 (CH, C-5 Thiazole); 118.66 (CH, Ar); 115.58
(2C, CH, Ar); 72.05 (CH2); 37.7 (CH2); 19.02 (C4thiazoleeCH3). Anal.
Calcd. (%) for C31H23F3N4O3S2 (620.66): C, 59.99; H, 3.74; N, 9.03; S,
10.33. Found: C, 59.81; H, 3.74; N, 9.01; S, 10.29. MS (EI, 70 eV): m/z
621 (Mþ), 352 [M- C12H7SOF3], 335 [M- C12H8NSOF3], 217
[C12H10NSO].
6.1.1.3. (2-p-Tolyl-thiazol-4-yl)-acetic acid [4-(2-phenyl-thiazol-4-
ylmethoxy)-benzylidene]-hydrazide (10). Yield 82%. White powder,
mp: 170e3  C. IR (KBr, cm1): 3253 (NH), 3094 (CHarom), 2920 and
2849 (CHaliph), 1671 (C]O Hydrazide), 1615 (C]C), 1540 (C]N). 1H
NMR (DMSO-d6, 500 MHz, ppm): d 11.35 (s, 1H, NH); 8.198 (s, 1H,
eCH]); 7.972e7.953 (dd, 2H, Ar-H); 7.823e7.807 (m, 3H, Ar-H);
7.68e7.656 (dd, 2H, Ar-H); 7.511 (d, 2H, Ar-H); 7.485 (s, 1H, H-
Thiazole); 7.448 (s, 1H, H-Thiazole); 7.315e7.290 (dd, 2H, Ar-H);
7.170e7.126 (dd, 2H, Ar-H); 5.286 (s, 2H, eCH2e); 4.194 (s, 2H,
eCH2e); 2.37 (s, 3H, eCH3). 13C NMR (DMSO-d6, 125 MHz, ppm):
d 171.19 (C-2 Thiazole); 168.97 (C]O, hydrazide); 166.04 (C-2
Thiazole); 164.02 (C, Ar); 159.58 (C-4 Thiazole); 156.79 (C-4
531
Thiazole); 146.89 (eCH]N); 144.29 (C, Ar); 143.32 (C, Ar); 134.67
(C, Ar); 131.86 (2C, CH, Ar); 131.01 (C, Ar); 131.09 (2C, CH, Ar);
128.47 (2C, CH, Ar); 126.96 (2C, CH, Ar); 126.04 (CH, C-5 Thiazole);
122.43 (2C, CH, Ar); 119.29 (CH, C-5 Thiazole); 118.75 (CH, Ar);
115.59 (2C, CH, Ar); 65.90 (CH2); 37.79 (CH2); 21.41 (CH3). Anal.
Calcd. (%) for C29H24N4O2S2 (524.65): C, 66.39; H, 4.61; N, 10.68; S,
12.22. Found: C, 66.15; H, 4.6; N, 10.64; S, 12.19. MS (EI, 70 eV): m/z
524 [M], 174 [C10H8NS], 104 [C7H5O].
6.1.1.4. [2-(3-Trifluoromethyl-phenyl)-thiazol-4-yl]-acetic acid [4-(2-
phenyl-thiazol-4-ylmethoxy)-benzylidene]-hydrazide (11). Yield
79%. White powder, mp: 175e6  C. IR (KBr, cm1): 3442 (NH), 3063
(CHarom), 2922 and 2850 (CHaliph), 1672 (C]O Hydrazide), 1599
(C]C), 1524 (C]N). 1H NMR (DMSO-d6, 500 MHz, ppm): d 11.38 (s,
1H, NH); 8.205 (s, 1H, eCH]); 8.194 (s, 1H, Ar-H); 7.991e7.954 (m,
4H, Ar-H); 7.809 (d, 2H, Ar-H); 7.682e7.655 (m, 2H, Ar-H); 7.622 (s,
1H, H-Thiazole); 7.521 (s, 1H, H-Thiazole); 7.507 (d, 2H, Ar-H);
7.169e7.123 (dd, 2H, Ar-H); 5.28 (s, 2H, eCH2e); 4.24 (s, 2H,
eCH2e). 13C NMR (DMSO-d6, 125 MHz, ppm): d 171.19 (C-2 Thia-
zole); 168.06 (C-2 Thiazole); 165.44 (C]O, hydrazide); 165.11 (]
CeO, Ar); 164.65 (C-4 Thiazole); 160.03 (C-4 Thiazole); 153.15
(eCH]N); 152.20 (C, Ar); 146.98 (C, Ar); 143.30 (C, Ar); 134.39 (CH,
Ar); 133.34 (CH, Ar); 131.08 (2C, CH, Ar); 130.84 (C, Ar); 130.57 (2C,
CH, Ar); 129.74 (CH, Ar); 128.98 (2C, CH, Ar); 127.69 (CF3); 126.61
(CH, C-5 Thiazole); 122.38 (CH, Ar); 119.2 (CH, C-5 Thiazole); 118.69
(CH, Ar); 115.58 (2C, CH, Ar); 65.94 (CH2); 37.7 (CH2). Anal. Calcd.
(%) for C29H21F3N4O2S2 (578.62): C, 60.20; H, 3.66; N, 9.68; S, 11.08.
Found: C, 60.11; H, 3.65; N, 9.65; S, 11.11. MS (EI, 70 eV): m/z 578 [M],
243 [C11H7NSCF3], 174 [C10H8NS], 104 [C7H5O], 71 [C2HNS].
6.1.1.5. (2-p-Tolyl-thiazol-4-yl)-acetic acid [3-(2-phenyl-thiazol-4-
ylmethoxy)-benzylidene]-hydrazide (12). Yield 80%. White powder,
mp: 173  C. IR (KBr, cm1): 3253 (NH), 3094 (CHarom), 2920 and
2849 (CHaliph), 1671 (C]O Hydrazide), 1615 (C]C), 1540 (C]N). 1H
NMR (DMSO-d6, 500 MHz, ppm): d 11.34 (s, 1H, NH); 8.2 (s, 1H,
eCH]); 7.913e7.886 (m, 3H, Ar-H); 7.821e7.808 (m, 3H, Ar-H);
7.678e7.657 (dd, 2H, Ar-H); 7.482 (s, 1H, H-Thiazole); 7.442 (s, 1H,
H-Thiazole); 7.311e7.286 (dd, 2H, Ar-H); 7.22 (m, 1H, Ar-H);
7.171e7.128 (dd, 2H, Ar-H); 5.28 (s, 2H, eCH2e); 4.19 (s, 2H,
eCH2e); 2.39 (s, 3H, eCH3). 13C NMR (DMSO-d6, 125 MHz, ppm):
d 171.52 (C-2 Thiazole); 165.66 (C]O, hydrazide); 165.43 (C-2
Thiazole); 165.01 (]CeO, Ar); 164.87 (C-4 Thiazole); 158.99 (C-4
Thiazole); 151.90 (eCH]N); 146.69 (C, Ar); 143.26 (C, Ar); 134. 58
(C, Ar); 133.36 (C, Ar); 131.78 (2C, CH, Ar); 130.62 (CH, Ar); 130.43
(2C, CH, Ar); 128.71 (2C, CH, Ar); 122.37 (CH, C-5 Thiazole); 119.01
(CH, Ar); 118.85 (CH, C-5 Thiazole); 126.62 (2C, CH, Ar); 118.67 (CH,
Ar); 117.19 (CH, Ar); 112.60 (CH, Ar); 65.94 (CH2); 37.71 (CH2); 21.40
(CH3). Anal. Calcd. (%) for C29H24N4O2S2 (524.65): C, 66.39; H, 4.61;
N, 10.68; S, 12.22. Found: C, 66.63; H, 4.6; N, 10.64; S, 12.19. MS (EI,
70 eV): m/z 524 [M], 247 [C6H5N3O], 232 [C12H11N2OS], 215
[C12H10NOS], 189 [C11H10NS], 174 [C10H8NS], 104 [C7H5O], 71
[C2HNS].
6.1.1.6. [2-(3-Trifluoromethyl-phenyl)-thiazol-4-yl]-acetic acid [3-(2-
phenyl-thiazol-4-ylmethoxy)-benzylidene]-hydrazide (13). Yield
72%. White powder, mp: 187e9  C. IR (KBr, cm1): 3445 (NH), 3065
(CHarom), 2920 and 2852 (CHaliph), 1675 (C]O Hydrazide), 1600
(C]C), 1521 (C]N). 1H NMR (DMSO-d6, 500 MHz, ppm): d 11.32 (s,
1H, NH); 8.2 (s, 1H, eCH]); 8.178 (s, 1H, Ar-H); 7.989e7.948 (m, 4H,
Ar-H); 7.779e7.769 (m, 3H, Ar-H); 7.68 (d, 2H, Ar-H); 7.62 (s, 1H, H-
Thiazole); 7.523 (s, 1H, H-Thiazole); 7.51e7.432 (m, 2H, Ar-H);
7.165e7.151 (m, 1H, Ar-H); 5.22 (s, 2H, eCH2e); 4.2 (s, 2H, eCH2e).
13
C NMR (DMSO-d6, 125 MHz, ppm): d 171.48 (C-2 Thiazole); 167.88
(C-2 Thiazole); 165.68 (C]O, hydrazide); 165.07 (]CeO, Ar);
164.67 (C-4 Thiazole); 158.92 (C-4 Thiazole); 151.95 (eCH]N);
532 C.M. Moldovan et al. / European Journal of Medicinal Chemistry 46 (2011) 526e534
147.01 (C, Ar); 143.28 (C, Ar); 134.39 (C, Ar); 133.36 (C, Ar); 131.12
(CH, Ar); 130.83 (CH, Ar); 130.60 (CH, Ar); 130.45 (2C, CH, Ar);
129.72 (CH, Ar); 126.60 (2C, CH, Ar); 122.38 (CH, C-5 Thiazole);
120.50 (CH, Ar); 119 (CH, Ar); 118.86 (CH, C-5 Thiazole); 118.65 (CH,
Ar); 117.15 (CH, Ar); 113.30 (CF3); 112.62 (CH, Ar); 65.94 (CH2); 37.71
(CH2). Anal. Calcd. (%) for C29H21F3N4O2S2 (578.62): C, 60.20; H,
3.66; N, 9.68; S, 11.08. Found: C, 60.40; H, 3.65; N, 9.66; S, 11.03. MS
(EI, 70 eV): m/z 578 [M], 336 [M- C11H7F3NS], 270 [C12H7F3NOS],
243 [C11H7F3NS], 174 [C10H8NS], 104 [C7H5O].
6.1.1.7. (2-p-Tolyl-thiazol-4-yl)-acetic acid {2-[2-(4-bromo-phenyl)-
thiazol-4-ylmethoxy]-benzylidene}-hydrazide (14). Yield 68%. White
powder, mp: 221e3  C. IR (KBr, cm1): 3445 (NH), 3087 (CHarom),
2921 and 2839 (CHaliph), 1675 (C]O Hydrazide), 1621 (C]C), 1555
(C]N), 1072 (CeBr). 1H NMR (500 MHz, DMSO-d6, ppm): d 11.95 (s,
1H, NH); 8.3 (s, 1H, eCH]); 7.683e7.675 (dd, 2H, Ar-H); 7.52 (s, 1H,
H-Thiazole); 7.517e7.502 (m, 1H, Ar-H); 7.478e7.454 (dd, 2H, Ar-H);
7.42 (s, 1H, H-Thiazole); 7.378e7.356 (m, 3H, Ar-H); 7.24e7.22 (dd,
2H, Ar-H); 7.154e7.126 (dd, 2H, Ar-H); 5.27 (s, 2H, eCH2e); 4.2 (s,
2H, eCH2e); 2.38 (s, 3H, eCH3). 13C NMR (DMSO-d6, 125 MHz,
ppm): d 171.27 (C-2 Thiazole); 170.65 (C]O, hydrazide); 165.53 (C-
2 Thiazole); 154.69 (C-4 Thiazole); 151.76 (C-4 Thiazole); 150.98 (C,
Ar); 150.37 (eCH]N); 143.02 (C, Ar); 140.19 (C, Ar); 132.87 (C, Ar);
131.04 (2C, CH, Ar); 130.89 (CH, Ar); 130.18 (2C, CH, Ar); 128.91 (2C,
CH, Ar); 127.24 (CH, Ar); 126.48 (2C, CH, Ar); 124.15 (CareBr); 119.96
(CH, C-5 Thiazole); 119.82 (C, Ar); 117.01 (CH, C-5 Thiazole); 114.92
(CH, Ar); 110.87 (CH, Ar); 65.90 (CH2); 38.85 (CH2); 21.40 (CH3).
Anal. Calcd. (%) for C29H23BrN4O2S2 (603.55): C, 57.71; H, 3.84; N,
9.28; S, 10.62. Found: C, 57.51; H, 3.83; N, 9.26; S, 10.59. MS (EI,
70 eV): m/z 604 [Mþ1], 373 [M- C6H5N2O], 254 [BrC10H7NS], 189
[C11H10NS].
6.1.1.8. [2-(3-Trifluoromethyl-phenyl)-thiazol-4-yl]-acetic acid {2-[2-
(4-bromo-phenyl)-thiazol-4-ylmethoxy]-benzylidene}-hydrazide
(15). Yield 64%. White powder, mp: 218e9  C. IR (KBr, cm1): 3440
(NH), 3079 (CHarom), 2919 and 2847 (CHaliph), 1674 (C]O Hydra-
zide), 1615 (C]C), 1550 (C]N), 1070 (CeBr). 1H NMR (500 MHz,
DMSO-d6, ppm): d 11.97 (s, 1H, NH); 8.3 (s, 1H, eCH]);
7.986e7.812 (m, 2H, Ar-H); 7.688e7.663 (m, 3H, Ar-H); 7.526 (s, 1H,
H-Thiazole); 7.498e7.485 (dd, 1H, Ar-H); 7.479e7.461 (m, 3H, Ar-
H); 7.44 (s, 1H, H-Thiazole); 7.184e7.163 (dd, 1H, Ar-H); 7.148e7.132
(dd, 2H, Ar-H); 5.3 (s, 2H,eCH2); 4.23 (s, 2H, eCH2e). 13C NMR
(DMSO-d6, 125 MHz, ppm): d 171.20 (C-2 Thiazole); 170.74 (C]O,
hydrazide); 165.62 (C-2 Thiazole); 155.43 (C-4 Thiazole); 153.12 (C-
4 Thiazole); 152.21 (C, Ar); 152.01 (-CH]N); 143.20 (C, Ar); 135.37
(C, Ar); 133.36 (C, Ar); 131.64 (2C, CH, Ar); 130.89 (CH, Ar); 130.26
(CH, Ar); 129.27 (CH, Ar); 127.69 (CF3); 127.24 (CH, Ar); 127.18 (CH,
Ar); 126.50 (2C, CH, Ar); 124.82 (CareBr); 120.45 (CH, C-5 Thiazole);
120.36 (C, Ar); 119.76 (CH, Ar); 117.88 (CH, C-5 Thiazole); 114.90
(CH, Ar); 110.89 (CH, Ar); 65.90 (CH2); 39.11 (CH2). Anal. Calcd. (%)
for C29H20BrF3N4O2S2 (657.52): C 52.97, H 3.07, N 8.52, S 9.75.
Found: C 53.10, H 3.08, N 8.532, S 9.73. MS (EI, 70 eV): m/z 658
[Mþ1], 404 [BrC10H7NS], 373 [BrC17H12N2S], 358 [BrC17H12NS], 269
[BrC10H7NSO], 254 [BrC10H7NS].
6.1.1.9. [2-(3-Trifluoromethyl-phenyl)-thiazol-4-yl]-acetic acid {3-[2-
(4-bromo-phenyl)-thiazol-4-ylmethoxy]-benzylidene}-hydrazide
(16). Yield 73%. White powder, mp: 192e7  C. IR (KBr, cm1): 3445
(NH), 3075 (CHarom), 2921 and 2857 (CHaliph), 1675 (C]O Hydra-
zide), 1620 (C]C), 1508 (C]N), 1071 (CeBr). 1H NMR (500 MHz,
DMSO-d6, ppm): d 11.99 (s, 1H, NH); 8.2 (s, 1H, eCH]);
7.998e7.968 (m, 2H, Ar-H); 7.784e7.727 (m, 2H, Ar-H); 7.687e7.665
(dd, 2H, Ar-H); 7.526 (s, 1H, H-Thiazole); 7.498e7.488 (m, 1H, Ar-H);
7.478e7.465 (m, 2H, Ar-H); 7.44 (s, 1H, H-Thiazole); 7.139e7.122 (m,
3H, Ar-H); 5.28 (s, 2H, eCH2e); 4.17 (s, 2H, eCH2e). 13C NMR
(DMSO-d6, 125 MHz, ppm): d 171.25 (C-2 Thiazole); 167.12 (C-2
Thiazole); 166.67 (C]O, hydrazide); 165.53 (]CeO, Ar); 160.28 (C-
4 Thiazole); 153.32 (C-4 Thiazole); 151.65 (eCH]N); 146.91 (C, Ar);
143.22 (C, Ar); 143.45 (C, Ar); 140.34 (2C, CH, Ar); 133.97 (C, Ar);
131.12 (CH, Ar); 130.83 (CH, Ar); 130.60 (CH, Ar); 129.61 (CH, Ar);
126.53 (2C, CH, Ar); 124.35 (CareBr); 122.57 (CH, C-5 Thiazole);
120.52 (CH, Ar); 119.22 (CH, Ar); 118.88 (CH, C-5 Thiazole); 118.68
(CH, Ar); 113.28 (CF3); 112.58 (CH, Ar); 65.87 (CH2); 37.70 (CH2).
Anal. Calcd. (%) for C29H20BrF3N4O2S2 (657.52): C, 52.97; H, 3.07; N,
8.52; S, 9.75. Found: C, 53.99; H, 3.08; N, 8.53; S, 9.74. MS (EI,
70 eV): m/z 657 [M], 417 [M-C11H7NSF3], 404 [M-C10H7NSBr], 254
[BrC10H7NS], 71 [C2HNS].
6.1.1.10. (2-p-Tolyl-thiazol-4-yl)-acetic acid {4-[2-(4-bromo-phenyl)-
thiazol-4-ylmethoxy]-benzylidene}-hydrazide (17). Yield 89.4%.
Yellow powder, mp: 204e5  C. IR (KBr, cm1): 3445 (NH), 3079
(CHarom), 2921 and 2837 (CHaliph), 1674 (C]O Hydrazide), 1621 (C]
C), 1514 (C]N), 1070 (CeBr). 1H NMR (500 MHz, DMSO-d6, ppm):
d 11 (s, 1H, NH); 8.78 (d, 2H, Ar-H); 8.41 (s, 1H, eCH]); 8.04 (dd, 2H,
Ar-H); 7.82 (dd, 2H, Ar-H); 7.72 (dd, 2H, Ar-H); 7.54 (dd, 2H, Ar-H);
7.52 (s, 1H, H-Thiazole); 7.48 (s, 1H, H-Thiazole); 7.13 (dd, 2H, Ar-H);
5.32 (s, 2H, eCH2e); 4.2 (s, 2H, eCH2e); 2.38 (s, 3H, eCH3e). 13C
NMR (DMSO-d6, 125 MHz, ppm): d 171.20 (C-2 Thiazole); 168.88
(C]O, hydrazide); 165.53 (C-2 Thiazole); 159.99 (]CeO, Ar);
153.31 (C-4 Thiazole); 151.63 (C-4 Thiazole); 146.89 (eCH]N);
143.22 (C, Ar); 140.39 (C, Ar); 132.62 (C, Ar); 130.99 (2C, CH, Ar);
130.21 (2C, CH, Ar); 128.99 (2C, CH, Ar); 128.54 (C, Ar); 127.73 (2C,
CH, Ar); 126.41 (2C, CH, Ar); 124.14 (CareBr); 119.78 (CH, C-5
Thiazole); 116.81 (CH, C-5 Thiazole); 115.60 (2C, CH, Ar); 65.85
(CH2); 37.83 (CH2); 21.39 (CH3). Anal. Calcd. (%) for C29H23BrN4O2S2
(603.55): C, 57.71; H, 3.84; N, 9.28; S, 10.62. Found: C, 57.55; H, 3.83;
N, 9.27; S, 10.59. MS (EI, 70 eV): m/z 604 [Mþ1], 373 [M- C6H5N2O],
254 [BrC10H7NS], 189 [C11H10NS].
6.1.1.11. [2-(3-Trifluoromethyl-phenyl)-thiazol-4-yl]-acetic acid {4-
[2-(4-bromo-phenyl)-thiazol-4-ylmethoxy]-benzylidene}-hydrazide
(18). Yield 81%. White powder, mp: 195e6  C. IR (KBr, cm1): 3442
(NH), 3077 (CHarom), 2920 and 2839 (CHaliph), 1675 (C]O Hydra-
zide), 1621 (C]C), 1520 (C]N), 1072 (CeBr). 1H NMR (500 MHz,
DMSO-d6, ppm): d 11.95 (s, 1H, NH); 8.4 (s, 1H, eCH]);
8.227e8.198 (m, 1H, Ar-H); 8.025e8.019 (m, 1H, Ar-H); 8.016 (dd,
2H, Ar-H); 7.864e7.847 (m, 2H, Ar-H); 7.718 (dd, 2H, Ar-H); 7.523
(dd, 2H, Ar-H); 7.516 (s, 1H, H-Thiazole); 7.447 (s, 1H, H-Thiazole);
7.121 (dd, 2H, Ar-H); 5.34 (s, 2H, eCH2); 4.21 (s, 2H, eCH2e). 13C
NMR (DMSO-d6, 125 MHz, ppm): d 171.25 (C-2 Thiazole); 168.88
(C]O, hydrazide); 165.59 (C-2 Thiazole); 160.03 (]CeO, Ar);
153.61 (C-4 Thiazole); 151.72 (C-4 Thiazole); 146.76 (eCH]N);
143.12 (C, Ar); 140.41 (C, Ar); 133.09 (2C, CH, Ar); 132.67 (C, Ar);
130.21 (CH, Ar); 129.81 (CH, Ar); 128.48 (C, Ar); 127.67 (2C, CH, Ar);
127.29 (CH, Ar); 126.47 (2C, CH, Ar); 124.11 (CareBr); 119.78 (CH, C-
5 Thiazole); 118.81 (CH, Ar); 116.84 (CH, C-5 Thiazole); 115.61 (2C,
CH, Ar); 114.25 (CF3); 65.89 (CH2); 37.82 (CH2). Anal. Calcd. (%) for
C29H20BrF3N4O2S2 (657.52): C, 52.97; H, 3.07; N, 8.52; S, 9.75.
Found: C, 53.00; H, 3.08; N, 8.54; S, 9.73. MS (EI, 70 eV): m/z 657
[M], 417 [M-C11H7NSF3], 404 [M-C10H7NSBr], 254 [BrC10H7NS], 71
[C2HNS].
6.1.1.12. (4-Oxo-2-phenyl-4H-chromen-7-yloxy)-acetic acid [4-(2-
phenyl-thiazol-4-ylmethoxy)-benzylidene]-hydrazide (20). Yield
80%. White powder, mp: 234e5  C. IR (KBr, cm1): 3446 (NH), 3123
(CHarom), 2926 (CHaliph), 1682 (C]O Chromone), 1650 (C]O
hydrazide), 1627 (C]C), 1522 (C]N). 1H NMR (DMSO-d6, 500 MHz,
ppm): d 11.59 (s, 1H, NH); 8.3 (s, 1H, eCH]); 7.729 (d, 2H, Ar-H);
7.685 (d, 2H, Ar-H); 7.605e7.599 (d, 1H, Ar-H); 7.585 (t, 1H, Ar-H);
 C.M. Moldovan et al. / European Journal of Medicinal Chemistry 46 (2011) 526e534
7.568 (d, 2H, Ar-H); 7.526 (s, 1H, H-Thiazole); 7.513 (d, 2H, Ar-H);
7.379 (d, 1H, Ar-H); 7.348 (d, 2H, Ar-H); 7.180e7.163 (m, 2H, Ar-H);
7.143e7.121 (dd, 2H, Ar-H); 6.976 (s, 1H, Ar-H); 5.29 (s, 2H, eCH2e);
4.87 (s, 2H, eCH2e). 13C NMR (DMSO-d6, 125 MHz, ppm): d 176.90
(C]O, chromone); 168.58 (C-2 Thiazole); 167.96 (C]O, hydrazide);
163.73 (C, Ar); 163.48 (C, Ar); 162.73 (C, Ar); 160.12 (C-4 Thiazole);
157.81 (C, Ar); 153.11 (eCH]N); 144.33 (C, Ar); 133.35 (C, Ar);
132.15 (CH, Ar); 131.65 (2C, CH, Ar); 130.89 (C, Ar); 129.78 (2C, CH,
Ar); 129.57 (2C, CH, Ar); 129.29 (CH, Ar); 129.11 (2C, CH, Ar); 127.42
(2C, CH, Ar); 126.67 (CH, C-5 Thiazole); 119.31 (C, Ar); 117.83 (CH,
Ar); 115.63 (2C, CH, Ar); 115.46 (CH, Ar); 107.23 (CH, Ar); 102.36
(CH, Ar); 65.95 (CH2), 65.88 (CH2). Anal. Calcd. (%) for C34H25N3O5S
(587.64): C, 69.49; H, 4.29; N, 7.15; S, 5.46. Found: C, 69.34; H, 4.28;
N, 7.14; S, 5.44. MS (EI, 70 eV): m/z 587 [M], 310 [M- C17H13NOS],
295 [C17H13N2OS], 278 [C17H13NOS], 174 [C10H8NS], 104 [C7H5O], 71
[C2HNS].
6.1.1.13. (4-Oxo-2-phenyl-4H-chromen-7-yloxy)-acetic acid [3-(2-
phenyl-thiazol-4-ylmethoxy)-benzylidene]-hydrazide (21). Yield
76%. White powder, mp: 202e4  C. IR (KBr, cm1): 3446 (NH),
3123 (CHarom), 2926 (CHaliph), 1683 (C]O Chromone), 1649 (C]O
Hydrazide), 1627 (C]C), 1522 (C]N). 1H NMR (DMSO-d6,
500 MHz, ppm): d 11.68 (s, 1H, NH); 8.42 (d, 1H, Ar-H); 8.24 (s, 1H,
eCH]); 8.128e8.104 (d, 2H, Ar-H); 7.976e7.962 (m, 3H, Ar-H);
7.742e7.721 (m, 2H, Ar-H); 7.678e7.649 (m, 4H, Ar-H); 7.536 (d,
2H, Ar-H); 7.522 (s, 1H, H-Thiazole); 7.139e7.119 (m, 3H, Ar-H);
6.965 (s, 1H, Ar-H); 5.24 (s, 2H, eCH2e); 4.81 (s, 2H, eCH2e). 13C
NMR (DMSO-d6, 125 MHz, ppm): d 176.92 (C]O, chromone);
168.59 (C-2 Thiazole); 167.91 (C]O, hydrazide); 163.82 (C, Ar);
163.71 (C, Ar); 163.28 (C, Ar); 160.72 (C-4 Thiazole); 157.82 (C,
Ar); 152.86 (eCH]N); 147.24 (C, Ar); 144.28 (C, Ar); 133.30 (C,
Ar); 132.20 (CH, Ar); 131.61 (2C, CH, Ar); 130.73 (CH, Ar); 129.69
(2C, CH, Ar); 129.21 (CH, Ar); 129.11 (2C, CH, Ar); 127.53 (2C, CH,
Ar); 126.49 (CH, C-5 Thiazole); 123.47 (CH, Ar); 120.04 (CH, Ar);
119.36 (C, Ar); 117.88 (CH, Ar); 115.61 (CH, Ar); 115.39 (CH, Ar);
107.29 (CH, Ar); 102.38 (CH, Ar); 65.93 (CH2); 65.87 (CH2). Anal.
Calcd. (%) for C34H25N3O5S (587.64): C, 69.49; H, 4.29; N, 7.15; S,
5.46. Found: C, 69.23; H, 4.28; N, 7.13; S, 5.47. MS (EI, 70 eV): m/z
587 [M], 238 [C15H9O3], 174 [C10H8NS], 104 [C7H5O].
6.1.1.14. (4-Oxo-2-phenyl-4H-chromen-7-yloxy)-acetic acid {3-[2-(4-
bromo-phenyl)-thiazol-4-ylmethoxy]-benzylidene}-hydrazide
(22). Yield 77%. White powder, mp: 216e8  C. IR (KBr, cm1): 3445
(NH), 3075 (CHarom), 2921 and 2857 (CHaliph), 1685 (C]O Chro-
mone), 1652 (C]O Hydrazide), 1620 (C]C), 1541 (C]N), 1071
(CeBr). 1H NMR (500 MHz, DMSO-d6, ppm): d 12.1 (s, 1H, NH); 8.43
(d, 1H, Ar-H); 8.2 (s, 1H, eCH]); 8.129e8.115 (dd, 2H, Ar-H);
7.994e7.978 (m, 3H, Ar-H); 7.763e7.745 (m, 2H, Ar-H); 7.685e7.66
(dd, 2H, Ar-H); 7.627e7.615 (dd, 1H, Ar-H); 7.422 (s, 1H, H-Thia-
zole); 7.257e7.244 (m, 3H, Ar-H); 7.238e7.22 (m, 2H, Ar-H); 7.12 (s,
1H, Ar-H); 5.4 (s, 2H, eCH2e); 4.37 (s, 2H, eCH2e). 13C NMR
(DMSO-d6, 125 MHz, ppm): d 176.93 (C]O, chromone); 168.64 (C-2
Thiazole); 168.03 (C]O, hydrazide); 163.91 (C, Ar); 163.68 (C, Ar);
163.22 (C, Ar); 160.70 (C-4 Thiazole); 157.75 (C, Ar); 152.73 (-CH]
N); 147.21 (C, Ar); 144.17 (C, Ar); 133.45 (2C, CH, Ar); 133.30 (C, Ar);
132.20 (CH, Ar); 131.61 (2C, CH, Ar); 130.73 (CH, Ar); 129.22 (CH,
Ar); 129.19 (2C, CH, Ar); 127.50 (2C, CH, Ar); 127.03 (CareBr); 126.42
(CH, C-5 Thiazole); 123.39 (CH, Ar); 120.11 (CH, Ar); 119.32 (C, Ar);
115.58 (CH, Ar); 115.27 (CH, Ar); 107.30 (CH, Ar); 102.40 (CH, Ar);
65.94 (CH2); 65.86 (CH2). Anal. Calcd. (%) for C34H24BrN3O5S
(666.54): C, 61.27; H, 3.63; N, 6.3; S, 4.81. Found: C, 61.32; H, 3.62; N,
6.28; S, 4.8. MS (EI, 70 eV): m/z 667 [Mþ1], 372 [C17H12N2SBr], 295
[C17H12NO4], 254 [C10H7NSBr], 238 [C15H9O3], 182 [C7H4NBr], 71
[C2HNS].
533
6.2. Pharmacology
6.2.1. Animals
The experiments were performed on adult male Wistar-Bra-
tislava albino rats, weighing 200e250 g. The animals were obtained
from the Biobase of University of Medicine and Pharmacy Cluj-
Napoca and housed at 25  2  C, 50  5% relative humidity and 12 h
light/dark cycle. They were distributed in groups of ten and had free
access to water and food. All the experimental procedures and
protocols used in this study were reviewed and approved by the
Institutional Animal Ethical Committee (IAEC) of University of
Medicine and Pharmacy Cluj-Napoca. Experiments were performed
in triplicate.
6.2.2. Anti-inflammatory activity
For the group called Inflammation, each animal was injected i.m.
with 0.6 mL/100 g (body weight) of turpentine oil, the pro-
inflammatory substance. The same procedure and dose were used
for the other groups, too. After that, a 3.2 mg/kg dose, equivalent to
0.0091168 mmol/kg of Meloxicam, the reference standard drug,
was administered i.p. to the animals from the reference group. The
test groups received the synthesized compounds in an equi-molar
dose with Meloxicam, by the i.p. administration of its 1% carbox-
ymethyl celullose suspension.
6.2.2.1. In vitro phagocytosis test. Investigation of phagocytic
activity of PMN cell was performed as described in literature
[31e40] with slight modifications. Blood samples from treated
animals were incubated in siliconated tube with E. coli suspension
at 37  C for 30 min. Smears were prepared on slides for microscopy.
Cells were fixed with methanol, stained with May-Grünwald
Giemsa and examined microscopically. Phagocytic activity was
assessed by calculating two parameters: the phagocytic index (PI)
(PI% ¼ phagocytes with at least one phagocyted germ from 200
leukocytes counted) and the phagocytic activity (PA) (PA ¼ number
of germs phagocyted by 100 leukocytes).
6.2.2.2. Measurement of serum nitrate and nitrite. Reduction of
nitrate to nitrite occurs in acidic solution by adding VCl3 (100 ml) to
serum samples (100 ml), rapidly followed by addition of the Griess
reagents, SULF (50 ml) and NEDD (50 ml). The absorbance at
540 nm was measured following incubation (usually 30e45 min).
6.2.3. Statistical analysis
The values are expressed as mean  S.D. for Inflammation
group, Meloxicam group and the healthy population, separately.
The comparisons of parameters were performed with Student’s
t-test and correlation analyses were performed using Spearman
correlation test. A p-value <0.05 was accepted as significant. Data
were analyzed using the SPSS for Windows computing program
(Version 11.0).
Acknowledgements
The authors would like to thank The National University
Research Council in Romania (scholarship BD109/2007) for the
financial support.
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